Detection of amplified nucleic acid sequences using bifunctional haptenization and dyed microparticles
First Claim
1. A method for the detection of amplified target nucleic acid sequences which comprises:
- a) denaturing a multiple stranded nucleic acid to produce a single stranded nucleic acid or beginning with a single stranded nucleic acid;
b) amplifying the target nucleic acid sequence with a first hapten derivatized complementary primer and dNTP and a second hapten derivatized complementary primer and dNTP to produce bifinctionally labeled specific target nucleic acid amplicons;
c) contacting dyed microparticles conjugated to a first hapten specific receptor corresponding to said first primer, dyed microparticles conjugated to a second hapten specific receptor corresponding to said second primer and said labeled target nucleic acid sequence wherein an agglutination reaction forms agglutination product of the amplified target nucleic acid; and
d) detecting the resulting agglutination product.
8 Assignments
0 Petitions
Accused Products
Abstract
The invention describes an assay for detecting amplified target nucleic acid sequences with a visual signal. The sensitivity and specificity of the methodology are based on bifunctional target labeling during the amplification step or subsequent hybridization that generates a bifunctional label. The invention may be used, for example, in the screening of amplicon detection for the purpose of more efficiently screening libraries. The invention is also useful to detect nucleic acid sequences indicative of a genetic defect or contagious disease when used with the appropriate primers, as well as detect the existence of nucleic acid amplification.
100 Citations
30 Claims
-
1. A method for the detection of amplified target nucleic acid sequences which comprises:
-
a) denaturing a multiple stranded nucleic acid to produce a single stranded nucleic acid or beginning with a single stranded nucleic acid; b) amplifying the target nucleic acid sequence with a first hapten derivatized complementary primer and dNTP and a second hapten derivatized complementary primer and dNTP to produce bifinctionally labeled specific target nucleic acid amplicons; c) contacting dyed microparticles conjugated to a first hapten specific receptor corresponding to said first primer, dyed microparticles conjugated to a second hapten specific receptor corresponding to said second primer and said labeled target nucleic acid sequence wherein an agglutination reaction forms agglutination product of the amplified target nucleic acid; and d) detecting the resulting agglutination product. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25)
-
-
26. A kit for the amplification and detection of a target nucleic acid consisting essentially of:
-
a) target specific hapten labeled primers; b) thermostable polymerase; c) dNTPs; and d) receptor bound dyed microparticles. - View Dependent Claims (27, 28, 29, 30)
-
Specification